OBJECTIVE To investigate the relationship of the EphA2 gene with the occurrence, invasion and metastasis of esophageal carcinoma. METHODS The expression of EphA2 mRNA was detected by RT-PCR and the EphA2 protein was e...OBJECTIVE To investigate the relationship of the EphA2 gene with the occurrence, invasion and metastasis of esophageal carcinoma. METHODS The expression of EphA2 mRNA was detected by RT-PCR and the EphA2 protein was estimated by immunohistochemistry (SP method) in both esophageal cancerous tissues and normal epithelial tissues. RESULTS The expression of EphA2 mRNA showed no difference between esophageal cancerous tissues and normal epithelium, and there appeared to be no correlation with differentiation of the cancerous tissues, the depth of infiltration or lymph node metastasis (P>0.05). However, the expression of the EphA2 protein was significantly higher in cancerous tissues compared to normal epithelial tissues (P<0.05). The expression of the EphA2 protein in a deeper invasive group and in a group with lymph node metastasis was significantly higher compared to a superficially invasive group and a group without lymph node metastasis (P<0.05). Its expression did not appear to be correlated with differentiation of cancerous tissues (P>0.05). CONCLUSION The occurrence of esophagus carcinoma and the formation of invasion and metastasis may be related to overexpression of the EphA2 protein but not to the level of mRNA, a finding which may due to up-regulation at the translation level or by increased protein stability.展开更多
AIM:To identify the genetic defect in a Chinese family with bilateral progressive childhood posterior cataract. METHODS:A two-generation family was recruited in this study. Family history and clinical data were record...AIM:To identify the genetic defect in a Chinese family with bilateral progressive childhood posterior cataract. METHODS:A two-generation family was recruited in this study. Family history and clinical data were recorded. All reported candidate genes associated with congenital posterior cataract were screened by direct DNA sequencing. ·RESULTS:All affected individuals presented posterior opacities in the lens. Direct sequencing of the candidate genes showed a heterozygous c. 2668C 】T variation in EPHA2 gene, which resulted in the replacement of arginine by cysteine at codon 890 (p. R890C). This mutation was found in two affected individuals, but was not observed in 200 normal controls. ·CONCLUSION:We report a novel mutation (p. R890C) in the EPHA2 receptor tyrosine kinase gene. The finding expands the mutation spectrum of EPHA2 in association with posterior cataract.展开更多
Objective To establish a method cloning the exon 3 of eck gene from normal tissue and ZR 75 1 cell line (a human breast cancer cell line)and study whether these genes exist mutant. Methods Designed a pair of s...Objective To establish a method cloning the exon 3 of eck gene from normal tissue and ZR 75 1 cell line (a human breast cancer cell line)and study whether these genes exist mutant. Methods Designed a pair of specific primers and amplified the exon 3 of eck gene fragment from the extracted genomic DNA derived from normal epithelial cells from skin tissue and ZR 75 1 cell line respectively by PCR technique. Transformed the E.coil. JM109 with recombinant plamids constructed by inserting the amplified fragments into medium vector pUCm T and sequenced these amplified fragments after primary screening of endonuclease restriction digestion and PCR amplification. Results ① Obtained the genomic DNA of human normal epithelial cells and ZR 75 1 cell line respectively. ② Obtained the amplified fragments of human exon 3 of eck gene through PCR technique. ③ Obtained the cloning vectors of exon 3 of eck gene of human normal epithelial cells and ZR 75 1 cell line respectively. ④ ZR 75 1 cell line exists mutation of nucleotides. Conclusion Successfully established the method of cloning the human exon 3 of eck gene and found some mutations in the detected samples. This study lays a foundation for further studying the function of eck gene in tumorgenesis.展开更多
文摘OBJECTIVE To investigate the relationship of the EphA2 gene with the occurrence, invasion and metastasis of esophageal carcinoma. METHODS The expression of EphA2 mRNA was detected by RT-PCR and the EphA2 protein was estimated by immunohistochemistry (SP method) in both esophageal cancerous tissues and normal epithelial tissues. RESULTS The expression of EphA2 mRNA showed no difference between esophageal cancerous tissues and normal epithelium, and there appeared to be no correlation with differentiation of the cancerous tissues, the depth of infiltration or lymph node metastasis (P>0.05). However, the expression of the EphA2 protein was significantly higher in cancerous tissues compared to normal epithelial tissues (P<0.05). The expression of the EphA2 protein in a deeper invasive group and in a group with lymph node metastasis was significantly higher compared to a superficially invasive group and a group without lymph node metastasis (P<0.05). Its expression did not appear to be correlated with differentiation of cancerous tissues (P>0.05). CONCLUSION The occurrence of esophagus carcinoma and the formation of invasion and metastasis may be related to overexpression of the EphA2 protein but not to the level of mRNA, a finding which may due to up-regulation at the translation level or by increased protein stability.
基金Science and Technology Program of Zhejiang University, China (No. 2011FZA70130)Medical Science and Technology Project of Zhejiang Province, China(No. 2010QNA012)+1 种基金Zhejiang Key Innovation Team Project of China (No. 2009R50039)Zhejiang Key Laboratory Found of China (No. 2011E10006)
文摘AIM:To identify the genetic defect in a Chinese family with bilateral progressive childhood posterior cataract. METHODS:A two-generation family was recruited in this study. Family history and clinical data were recorded. All reported candidate genes associated with congenital posterior cataract were screened by direct DNA sequencing. ·RESULTS:All affected individuals presented posterior opacities in the lens. Direct sequencing of the candidate genes showed a heterozygous c. 2668C 】T variation in EPHA2 gene, which resulted in the replacement of arginine by cysteine at codon 890 (p. R890C). This mutation was found in two affected individuals, but was not observed in 200 normal controls. ·CONCLUSION:We report a novel mutation (p. R890C) in the EPHA2 receptor tyrosine kinase gene. The finding expands the mutation spectrum of EPHA2 in association with posterior cataract.
文摘Objective To establish a method cloning the exon 3 of eck gene from normal tissue and ZR 75 1 cell line (a human breast cancer cell line)and study whether these genes exist mutant. Methods Designed a pair of specific primers and amplified the exon 3 of eck gene fragment from the extracted genomic DNA derived from normal epithelial cells from skin tissue and ZR 75 1 cell line respectively by PCR technique. Transformed the E.coil. JM109 with recombinant plamids constructed by inserting the amplified fragments into medium vector pUCm T and sequenced these amplified fragments after primary screening of endonuclease restriction digestion and PCR amplification. Results ① Obtained the genomic DNA of human normal epithelial cells and ZR 75 1 cell line respectively. ② Obtained the amplified fragments of human exon 3 of eck gene through PCR technique. ③ Obtained the cloning vectors of exon 3 of eck gene of human normal epithelial cells and ZR 75 1 cell line respectively. ④ ZR 75 1 cell line exists mutation of nucleotides. Conclusion Successfully established the method of cloning the human exon 3 of eck gene and found some mutations in the detected samples. This study lays a foundation for further studying the function of eck gene in tumorgenesis.
